Leukocyte Adherence Inhibition Research Articles

EFFECTS OF SINUSOIDAL MAGNETIC FIELD ON ADHERENCE INHIBITION OF LEUKOCYTES

Response of leukocytes to exposure to an external magnetic field with frequency 50 Hz and sinusoidal waveform was investigated in vitro using the leukocyte adherence inhibition (LAI) assay developed as a measure of cell-mediated immunity. Leukocytes taken from healthy humans adhere, but their adherence decreases after 1 hr of exposure to the magnetic field with magnetic induction of 1 and 10 mT. The majority of leukocytes taken from cancer patients before any medical treatment do not adhere, and exposure to the magnetic field increases adherence. Correlation between the LAI assay results and the cell-mediated immunity suggests an effect of magnetic fields on leukocyte immune function in humans.

EVALUATION OF HEMATOLOGICAL AND IMMUNOLOGICAL PARAMETERS IN MICE EXPOSED TO SUBLETHAL AND SUBCHRONIC DOSES OF DIMETHOATE

Effects of sublethal and subchronic doses of dimethoate (DM), an organophosphorus pesticide, have been studied on hematological and immunological parameters in mice. The results revealed that DM-treated mice showed significant increase in red blood cells (RBC), white blood cells, percent monocytes, and percent basophils, whereas the percent neutrophils was significantly decreased.The antibody titres were higher on day 4 postantigen(sheep RBC/bovine serum albumin [BSA]) inoculation, but suppressed on day 7 and 11 postantigen inoculation in DM-treated mice. However, the three cell-mediated immune parameters-leukocyte migration inhibition (LMI), leukocyte adherence inhibition (LAI), and nitroblue tetrazolium (NBT) reduction-were invariably depressed in DM-treated BSA-immunized mice, whereas the percent LMI and LAI were lower and NBT was higher in the DM-treated SRBC-immunized mice. Our results show that exposure to DM suppresses humoral and cell-mediated immune response, but its effect on macrophage function...

Evaluation of host resistance and non‐specific cell mediated immune responses in mice exposed to sublethal subchronic doses of Aldrin

The effect of sublethal subchronic dose of Aldrin, an organochlorine pesticide, has been studied on non‐specific cell mediated immune responses and host resistance to an infection. Aldrin treatment in mice could result in significant suppression of stimulation of leukocytes to PHA as measured by the leukocyte migration inhibition and leukocyte adherence inhibition test. Further, it leads to a significant increase in parasitaemia, increased mortality rate and decreased survival time in mice infected with Plasmodium berghei. Our results reveal that a sublethal subchronic dose of Aldrin may affect non‐specific stimulation of cells involved in immune reactions and increase the rate of parasitaemia.

The Leukocyte Adherence Inhibition Test (LAI) in Preoperative Diagnosis of Epithelial Ovarian Cancer

One hundred two women admitted for evaluation of an adenexal mass were tested prior to surgery by the computerized tube leukocyte adherence inhibition (LAI) assay. The test was positive in 24/35 (69%) patients, diagnosed after surgery as epithelial ovarian cancer while 50/67 (75%) of those diagnosed as benign masses were negative. Only 2/38 (5%) of normal control subjects showed a false-positive result. The LAI test described in this report showed a sensitivity of 88% and a specificity of 75% in stage I ovarian cancer; thus it may be useful for early diagnosis of ovarian malignancy.

Identification of a human cancer related organ-specific neoantigen.

Human cancers express organ-specific neoantigens (OSNs) which elicit specific cellular immune responses in the cancer patient, as demonstrated by leukocyte adherence inhibition (LAI), an in vitro immune response assay. A purified protein of MW 40,000 (p40) exhibiting OSN (colon specific) activity was cleaved into specific peptide fragments and their partial amino acid sequences determined. This information was used in the polymerase chain reaction (PCR) to obtain a 992 bp cDNA clone (PCR-992) from a human colon adenocarcinoma cell line (LS-180). By comparison of the predicted amino acid sequence of PCR-992 with the known sequence of p40 peptides, PCR-992 was shown to correspond to almost the entire coding region of p40. Nucleotide sequence analysis suggested that the protein was mycoplasmal in origin due to its high A+T content (76%) and the presence of five in frame TGA termination codons; at least two of the latter are actually read as tryptophan, a known feature of mycoplasma translation. We have confirmed this origin by direct isolation of a contaminating mycoplasma species from the LS-180 cell line and demonstration that it could be hybridized with the PCR-992 probe. Northern and PCR analysis of RNA preparations from the contaminated LS-180 cell line showed that p40 was part of the high affinity transport system operon of Mycoplasma hyorhinis (Dudler et al, EMBO J., 7: 3963-3970, 1988). Total protein lysates of Mycoplasma hyorhinis cultivated without animal cells could elicit positive LAI responses when incubated with cancer patient leukocytes but not with normal patient leukocytes. The organ-specific nature of the response was, however, not observed indicating that host cell-mycoplasmal interactions may play a role in determining the organ-specific nature of p40 seen with the LAI. The significance of these findings will be discussed in the context of previous thinking regarding the origin of OSNs.

Leukocyte Adherence Inhibition Test: Cell-Mediated Reactivity to Tumor Antigens in Colorectal and Lung Tumor Long Survivors

Cell-mediated immunity versus tumor antigens (cytosols) of the same histotype and site was evaluated by means of the leukocyte adherence inhibition (LAI) test in 44 colorectal adenocarcinoma and 17 lung squamous-cell carcinoma patients 3-10 years after surgical resection. LAI-positivity was observed in 17/44 (38.6%) and 13/17 (76.5%) patients, respectively, together with serum blocking factors in 7/17 (41.2%) compared to 0/13. These results indicate that a high percentage of long survivors retain an immunologic memory of the tumor antigen to which they have been exposed. The prognostic significance of serum blocking factors is less clear.

RNA-mediated transfer of cellular immunity to a synthetic env antigen of the human immunodeficiency virus (HIV-1)

A sheep was immunized with a synthetic peptide corresponding to amino acid residues 586-606 of the precursor envelope protein GP-160 of the HIV-1 including a conserved epitope region of the GP-41 transmembrane protein in the mature viral particles, referred to as SM 284 HIV-1 [1]. It is demonstrated that immune RNA extracted from the lymphoid organs of the immunized animal (SM 284 HIV-1 I-RNA) was able to transfer immune cellular reactivity to SM 284 HIV-1 in vitro to human and rabbit lymphocytes and in vivo to Cebus apella monkeys. The transfer was detected by the leukocyte adherence inhibition test (LAI) as an indicator of cellular reactivity. One of the most relevant results was the demonstration that SM 284 HIV-1 I-RNA was able to induce cellular immunological memory in vivo in monkeys. These results may be relevant to delineate a new alternative for immunomodulation against HIV infection.

Tumor size, leukocyte adherence inhibition and serum levels of tumor antigen in dogs with the canine transmissible venereal sarcoma.

Tumor antigen (TA) associated with the canine transmissible venereal sarcoma (CTVS) was detected in the sera of dogs bearing the tumor. Rabbit antisera specific for tumor antigen and 3 M KCl extracts of CTVS cells were used in both a competitive enzyme-linked immunosorbent assay (ELISA) and antigen-capture ELISA to quantify levels of circulating TA. In a study of 29 dogs bearing the transplanted CTVS, levels of circulating TA correlated positively with tumor volume. In a longitudinal study of four dogs receiving a transplant of 10(8) viable CTVS cells, circulating CTVS antigen was detected transiently 2 days after transplantation, while persistent levels of TA associated with increasing tumor volume were demonstrable 19-34 days after transplantation. In three of four tumor-bearing dogs, levels of serum TA correlated inversely with values obtained with peripheral blood leukocytes in the leukocyte adherence inhibition (LAI) assay; elevated levels of circulating TA found in dogs with large (greater than 7 cm3) tumors were associated with decreased LAI reactivity of peripheral blood leukocytes. TA could not be detected in sera 48-72 h after surgical removal of CTVS whereas LAI reactivity of peripheral blood leukocytes to CTVS antigen rebounded 1-3 weeks following tumor excision. Results of this study support the use of the competitive ELISA and LAI techniques in assessing levels of circulating tumor antigen, tumor burden and tumor-specific immunity.

Suppressor cells and soluble factors from the spleens of UV‐irradiated mice

Several indices of splenic reactivity were measured daily after ultraviolet (UV) irradiation of mice. Spleen weight increased to a maximum after 5 days. Suppressor cell activity, manifested in various assays, peaked slightly earlier. The ability of passively transferred spleen cells to suppress the induction of contact hypersensitivity (CHS) was maximal after 4 days, as was the production of soluble suppressor factors in culture. The factors were detected by suppression of CHS in vivo and leucocyte adherence inhibition (LAI) in vitro. These assays appeared to detect different factors: an I-J- factor active in CHS assays and an I-J+ factor active in LAI assays. The latter factor required CD4-, CD8+ lymphocytes for its production.

Distinctive molecular markers and biological activities in two tumour‐specific murine T suppressor factors

Murine suppressor factors (SF) in tumour-bearer serum (TBS) and tumour-bearer spleen cell culture supernatants (SCCS) were compared with regard to their molecular markers, cellular requirements and suppressive activities. Suppressor factors in SCCS bear the idiotype I-J, and markers recognized by B16G and anti-lipomodulin antibodies require Ly2+ I-J+ cells for their production and cells of the same phenotypes, for suppression to be manifested in leucocyte adherence inhibition (LAI) assays, are not immunoglobulin (Igh)-restricted and are active in both the afferent and efferent phases of the immune response. Suppressor factors from TBS resemble those from SCCS with regard to idiotype, I-J and lipomodulin-like markers, but in contrast, do not bind to B16G antibody, require Ly2+ but not I-J+ cells for suppression to be manifested in LAI assays, are Igh-restricted, and suppress delayed-type hypersensitivity (DTH) reactions only when injected in the afferent phase. These results show that the SF from the different sources in tumour-bearing mice are not identical; they may be different parts of a suppressor cascade, or they may belong to entirely separate suppressor cascades.

Modified humoral leukocyte adherence inhibition test for detection of lung cancer : Kubasova T, Horvath M, Szarvas I, Beke CS, Koteles GJ. ‘Frederic Joliot-Curie’, National Research Institute for Radiobiology and Radiohygiene, H-1775 Budapest. Oncology 1989;46:386-90

Modified humoral leukocyte adherence inhibition test for detection of lung cancer : Kubasova T, Horvath M, Szarvas I, Beke CS, Koteles GJ. ‘Frederic Joliot-Curie’, National Research Institute for Radiobiology and Radiohygiene, H-1775 Budapest. Oncology 1989;46:386-90

Comparison of T suppressor factors from tumour-bearing mice and mice immunized with a monoclonal anti-idiotypic antibody.

Certain dosage schedules of a monoclonal anti-idiotypic antibody (related to a murine bladder carcinoma) were found to induce suppressor factor production by syngeneic mice. This suppressor factor resembled the factor from tumour-bearing mice with respect to idiotype specificity, possession of molecular markers (reactive with anti-IJ and B16G antibodies) and production by Lyt2+IJ+ T cells in spleen cell cultures. The two factors differed with respect to Igh restriction in an in vitro assay (leucocyte adherence inhibition) and ability to suppress the induction of delayed-type hypersensitivity to tumour antigen.

The role of the leukocyte adherence inhibition (LAI), CA 19-9, and tissue polypeptide antigen (TPA) tests in the diagnosis of pancreatic cancer.

The leukocyte adherence inhibition (LAI) assay measures host cell-mediated tumor immunity. The original test used by Halliday and Miller was modified by substituting tissue extracts with purified tumor antigens (gastrointestinal cancer antigen [GICA]) with the aim of reducing false-positive results in the diagnosis of patients with early pancreatic cancer. By our modified technique, the LAI assay identified four of five pancreatic cancers at an early stage and gave only one false-positive result in 16, showing a sensitivity of 80% and a specificity of 94%. Later stages of the disease responded poorly on the test. These results were significantly better than those by the CA 19-9 and tissue polypeptide antigen (TPA) tests. Should our findings be confirmed in larger material over a longer follow-up period, the LAI test could be proposed as a useful tool in the early diagnosis of pancreatic cancer.

Immune function in malignant cervical neoplasia: A multiparameter analysis

International Journal of Gynecology & ObstetricsVolume 29, Issue 2 p. 200-200 Citations from the literature oncology Immune function in malignant cervical neoplasia: A multiparameter analysis P Balaram, P BalaramSearch for more papers by this authorMR Pillai, MR PillaiSearch for more papers by this authorTK Padmanabhan, TK PadmanabhanSearch for more papers by this authorT Abraham, T AbrahamSearch for more papers by this authorNK Hareendran, NK HareendranSearch for more papers by this authorMK Nair, MK NairSearch for more papers by this author P Balaram, P BalaramSearch for more papers by this authorMR Pillai, MR PillaiSearch for more papers by this authorTK Padmanabhan, TK PadmanabhanSearch for more papers by this authorT Abraham, T AbrahamSearch for more papers by this authorNK Hareendran, NK HareendranSearch for more papers by this authorMK Nair, MK NairSearch for more papers by this author First published: June 1989 https://doi.org/10.1016/0020-7292(89)90876-XAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume29, Issue2June 1989Pages 200-200 RelatedInformation

Protection against a syngeneic mammary adenocarcinoma by administration of idiotypic and anti-idiotypic antibodies.

A hybridoma secreting a monoclonal antibody (mAb) with specificity for tumor-associated cell surface antigens of a transplantable murine mammary adenocarcinoma (SMC-168) was prepared by fusion of syngeneic C3H/He spleen cells with SP2 myeloma cells. Mice which were pretreated with this mAb (C-73) were significantly resistant to the outgrowth of a tumorigenic dose of SMC-168 cells when compared to controls. The treated mice developed tumor-specific cell-mediated immunity, measured by leukocyte adherence inhibition (LAI), which was equal to that of mice immunized with live tumor cells. The IgG fraction from serum of mice receiving mAb C-73 contained antibodies which would bind to that mAb suggesting the presence of anti-idiotypic antibodies (anti-Id). This binding could be partially inhibited by a soluble 1-butanol cell surface extract of SMC-168. Rabbits were immunized with mAb C-73 to produce a polyclonal anti-Id. The purified and absorbed IgG fraction of this serum would bind only to mAb C-73 and not to other mAbs of the same isotype or normal C3H/HeN IgG. Binding of the rabbit anti-Id to mAb C-73 could be partially inhibited by soluble tumor-associated antigen extracted from SMC-168. Mice immunized with this polyclonal anti-Id vaccine developed tumor-specific cell-mediated immunity and were significantly resistant to the outgrowth of a tumorigenic dose of SMC-168.

The role of acupuncture in asthma: changes in airways dynamics and LTC4 induced LAI.

A cohort of nine extrinsic asthma patients were treated by means of acupuncture. Patients were followed up for changes in medical treatment, spirometry, skin reactivity to immediate type reactions, total serum IgE levels and reactivity of their leukocytes to leukotriene C4 challenge (LTC4 induced leukocyte adherence inhibition (LAI) assay). Our results show that after acupuncture, treated patients were able to reduce bronchodilator and taper completely corticosteroid therapy. No change in skin reactivity or in IgE levels were noted. However, acupuncture treatment was able to negate, in 66.6%, the positive LTC4 induced responses.

Modified Humoral Leukocyte Adherence Inhibition Test for Detection of Lung Cancer

The freshly separated indicator cells (suspension of leukocytes) used in humoral leukocyte adherence inhibition test were labeled either with 14C-amino acid mixture or 3H-concanavalin A (3H-ConA). Instead of counting the adherent cells, the amount of 'adherent' radioactivity was measured by a liquid scintillation spectrometer. By the modified method, sera from 25 lung-carcinoma-bearing patients as well as serum samples from 21 healthy persons were examined in the presence of crude antigens prepared from 'normal' lung tissue or lung tumors of various histologic types. Although the results demonstrated high specificity and reproducibility of both methods, the binding of 3H-ConA to the surface of adherent cells is more expressed and assures higher levels of radioactivity.

Comparison by leukocyte adherence inhibition of human immune response to cancer-associated immunogens, Thomsen-Friedenreich (T) and Tn, myelin basic protein, and organ-specific cancer neoantigens

Peripheral blood leukocytes from cancer patients exhibit nonadherence to glass as an index of antigen recognition when incubated individually with four distinct, soluble tumor-related substances. Crude cancer extracts, purified antigens, T and Tn, myelin basic protein (MBP), and organ-specific cancer neoantigens (OSN), all elicited narrow dose-dependent leukocyte adherence inhibition (LAI) response curves. The present study focused on the reasons for the narrow antigen dose-LAI response relationship. Between 9 and 20 pmol of antigens elicited the maximum number of nonadherent leukocytes; cleavage products of T antigen and the nonapeptide (T18) of MBP required about a 10-fold increase in molar concentration for the same LAI response. When crude cancer extracts were combined with pure antigen or the pure antigens were combined at concentrations shown to give maximum LAI responses, the positive LAI responses were negated. The chemoattractant LTB 4 at 10 −11 M triggered maximum LAI. But when MBP was added with the LTB 4 at progressively increasing concentrations, there was dose-dependent negation of LAI. The magnitude of LAI depended on the total amount of mediator released rather than the rate of release. When leukocytes from cancer patients were incubated with optimum to high concentrations of MBP, the supernatants contained a mediator that gave similar bell-shaped dose-LAI responses on control leukocytes indicating that leukocytes from cancer patients react to a much broader range of antigen concentration than indicated by the LAI assay alone. High antigen dose negated LAI because of excess mediator production. Antigen-generated mediators had a biphasic effect inducing nonadherence and then adherence of leukocytes.

Immune function in malignant cervical neoplasia: A multiparameter analysis

Immunological assessment was carried out in 67 patients with various stages and histological grades of squamous cell carcinoma of the uterine cervix and compared with normal controls to discover whether any of these parameters could be used as an indicator of the patient's clinical status. The results show a gross impairment of the various lymphocyte subpopulations and anti-tumor immunity. This impairment was more pronounced in patients with high tumor load and having a histology of poorly differentiated squamous cell carcinomas. None of the parameters studied (total leukocyte numbers, lymphocyte subpopulations, leukocyte adherence inhibition test, natural killer cell activity, circulating immune complexes, and blocking effect of autologous serum) could be used as a clinical indicator, since no correlation could be noted between immune impairment and either the lymphocyte subpopulations or the circulating immune complexes.

Production of suppressor factors induced by ultraviolet irradiation or cis-urocanic acid requires Lyt-2+ lymphocytes.

Both ultraviolet (UV) irradiation and cis-urocanic acid (UCA) are reported to be associated with the suppression of contact hypersensitivity (CHS) and the induction of a soluble factor which suppresses leukocyte adherence inhibition (LAI) assays in vitro. The cellular origin of the suppressor factor (SF) has now been investigated in vivo and in vitro, using monoclonal antibodies against lymphocyte surface markers to deplete certain cell populations, namely T lymphocytes bearing the L3T4 (helper) and Lyt-2 (suppressor) markers. Depletion of Lyt-2+ cells from irradiated mice in vivo and from spleen cell cultures in vitro led to the elimination of detectable levels of SF. Depletion of L3T4+ cells had no such effect. Similarly, Lyt-2+ cells (but not L3T4+ cells) were shown to be necessary for the production of SF by normal spleen cells cultured with cis-UCA. These data suggest that the production of SF following UV irradiation may be related to the action of cis-UCA on Lyg-2+ lymphocytes.